title: Comparison of commercially available target enrichment methods for next-generation sequencing.
abstract: Isolating high-priority segments of genomes greatly enhances the efficiency of next-generation sequencing (NGS) by allowing researchers to focus on their regions of interest. For the 2010-11 DNA Sequencing Research Group (DSRG) study, we compared outcomes from two leading companies, Agilent Technologies (Santa Clara, CA, USA) and Roche NimbleGen (Madison, WI, USA), which offer custom-targeted genomic enrichment methods. Both companies were provided with the same genomic sample and challenged to capture identical genomic locations for DNA NGS. The target region totaled 3.5 Mb and included 31 individual genes and a 2-Mb contiguous interval. Each company was asked to design its best assay, perform the capture in replicates, and return the captured material to the DSRG-participating laboratories. Sequencing was performed in two different laboratories on Genome Analyzer IIx systems (Illumina, San Diego, CA, USA). Sequencing data were analyzed for sensitivity, specificity, and coverage of the desired regions. The success of the enrichment was highly dependent on the design of the capture probes. Overall, coverage variability was higher for the Agilent samples. As variant discovery is the ultimate goal for a typical targeted sequencing project, we compared samples for their ability to sequence single-nucleotide polymorphisms (SNPs) as a test of the ability to capture both chromosomes from the sample. In the targeted regions, we detected 2546 SNPs with the NimbleGen samples and 2071 with Agilent's. When limited to the regions that both companies included as baits, the number of SNPs was ∼1000 for each, with Agilent and NimbleGen finding a small number of unique SNPs not found by the other.
authors.affiliation: Tufts University School of Medicine, Boston, Massachusetts 02452, USA. kip.bodi@tufts.edu
authors.forename: K
authors.initials: K
authors.lastname: Bodi
authors.forename: A G
authors.initials: AG
authors.lastname: Perera
authors.forename: P S
authors.initials: PS
authors.lastname: Adams
authors.forename: D
authors.initials: D
authors.lastname: Bintzler
authors.forename: K
authors.initials: K
authors.lastname: Dewar
authors.forename: D S
authors.initials: DS
authors.lastname: Grove
authors.forename: J
authors.initials: J
authors.lastname: Kieleczawa
authors.forename: R H
authors.initials: RH
authors.lastname: Lyons
authors.forename: T A
authors.initials: TA
authors.lastname: Neubert
authors.forename: A C
authors.initials: AC
authors.lastname: Noll
authors.forename: S
authors.initials: S
authors.lastname: Singh
authors.forename: R
authors.initials: R
authors.lastname: Steen
authors.forename: M
authors.initials: M
authors.lastname: Zianni
mesh_terms: D002875:Chromosomes / Q000235:genetics; D004247:DNA / Q000235:genetics*; D015894:Genome, Human; D005838:Genotype; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*
chemical_list: D004247:DNA
keywords: Agilent; Illumina; NimbleGen; targeted capture